Optimized skin optical clearing for optical coherence tomography monitoring of encapsulated drug delivery through the hair follicles

Hair follicles (HF) represent a drug delivery reservoir for improved treatment of skin disorders. Although various particulate systems play an important role in HF‐targeting, their optical monitoring in skin is challenging due to strong light scattering. Optical clearing is an effective approach allowing the increasing of particle detection depth in skin. The enhancement of optical probing depth (OPD) and optical detection depth (ODD) of particle localization using optical coherence tomography (OCT) was evaluated under application of various optical clearing agents (OCAs) together with skin permeability enhancers ex vivo in rats. Efficient OPD increasing was demonstrated for all investigated OCAs. However, skin dehydration under action of hyperosmotic agents led to the worsening of OCT‐contrast in dermis decreasing the ODD. Lipophilic agents provided optical clearing of epidermis without its dehydration. The highest ODD was obtained at application of a PEG‐400/oleic acid mixture. This OCA was tested in vivo showing beneficial ODD and OPD enhancement.     

An ssDNA virus infecting archaea: a new lineage of viruses with a membrane envelope

Archaeal organisms are generally known as diverse extremophiles, but they play a crucial role also in moderate environments. So far, only about 50 archaeal viruses have been described in some detail. Despite this, unusual viral morphotypes within this group have been reported. Interestingly, all isolated archaeal viruses have a double-stranded DNA (dsDNA) genome. To further characterize the diversity of archaeal viruses, we screened highly saline water samples for archaea and their viruses. Here, we describe a new haloarchaeal virus, Halorubrum pleomorphic virus 1 (HRPV-1) that was isolated from a solar saltern and infects an indigenous host belonging to the genus Halorubrum . Infection does not cause cell lysis, but slightly retards growth of the host and results in high replication of the virus. The sequenced genome (7048 nucleotides) of HRPV-1 is single-stranded DNA (ssDNA), which makes HRPV-1 the first characterized archaeal virus that does not have a dsDNA genome. In spite of this, similarities to another archaeal virus were observed. Two major structural proteins were recognized in protein analyses, and by lipid analyses it was shown that the virion contains a membrane. Electron microscopy studies indicate that the enveloped virion is pleomorphic (approximately 44 × 55 nm). HRPV-1 virion may represent commonly used virion architecture, and it seems that structure-based virus lineages may be extended to non-icosahedral viruses.     

Accumulation of phenolic compounds in birch leaves is changed by elevated carbon dioxide and ozone

Atmospheric change may affect plant phenolic compounds, which play an important part in plant survival. Therefore, we studied the impacts of CO₂ and O₃ on the accumulation of 27 phenolic compounds in the short‐shoot leaves of two European silver birch (Betula pendula Roth) clones (clones 4 and 80). Seven‐year‐old soil‐grown trees were exposed in open‐top chambers over three growing seasons to ambient and twice ambient CO₂ and O₃ concentrations singly and in combination in central Finland. Elevated CO₂ increased the concentration of the phenolic acids (+25%), myricetin glycosides (+18%), catechin derivatives (+13%) and soluble condensed tannins (+19%) by increasing their accumulation in the leaves of the silver birch trees, but decreased the flavone aglycons (?7%) by growth dilution. Elevated O₃ increased the concentration of 3,4′‐dihydroxypropiophenone 3‐β‐d ‐glucoside (+22%), chlorogenic acid (+19%) and flavone aglycons (+4%) by inducing their accumulation possibly as a response to increased oxidative stress in the leaf cells. Nevertheless, this induction of antioxidant phenolic compounds did not seem to protect the birch leaves from detrimental O₃ effects on leaf weight and area, but may have even exacerbated them. On the other hand, elevated CO₂ did seem to protect the leaves from elevated O₃ because all the O₃‐derived effects on the leaf phenolics and traits were prevented by elevated CO₂. The effects of the chamber and elevated CO₂ on some compounds changed over time in response to the changes in the leaf traits, which implies that the trees were acclimatizing to the altered environmental conditions. Although the two clones used possessed different composition and concentrations of phenolic compounds, which could be related to their different latitudinal origin and physiological characteristics, they responded similarly to the treatments. However, in some cases the variation in phenolic concentrations caused by genotype or chamber environment was much larger than the changes caused by either elevated CO₂ or O₃.     

Neural and mechanical responses of the triceps surae muscle group after 1 h of repeated fast passive stretches.

Experiments were carried out to examine interaction between mechanical changes of the muscle-tendon unit and reduced reflex sensitivity after repeated and prolonged passive muscle stretching (RPS). There is some evidence that this interaction might be relevant also during active stretch-shortening cycle type of fatigue tasks. The results demonstrated a clear deterioration of voluntarily and electrically induced muscle contractions after RPS. Maximal voluntary contraction (MVC), average electromyographic activities of the gastrocnemius and soleus muscles, and maximal twitch contraction decreased on average by 13.8, 10.4, 7.6, and 16.8%, respectively. In addition, there was a 14% lengthening in the total duration of the twitch. MVCs measured at different ankle joint angles revealed a downward and rightward shift in the torque-fascicle length curve after RPS. Interestingly, there was a crossing in the torque-fascicle length curves while measured at different activation levels but at the same joint angle before and after RPS. Even though no changes were observed in the activation level during MVCs, all the reflex parameters showed a clear reduction after RPS. This study presents evidence that repeated and prolonged passive muscle stretching can lead to some modification of material behavior of the aponeurosis-tendon system, such as stress relaxation and/or plastic deformation. In addition, altered material properties seem to affect proprioceptive feedback and, therefore, the motor unit activation in proportion to the contractile failure.     

Effect of copper intoxication on rat liver proteasome activity: relationship with oxidative stress

Copper toxicity is associated with formation of reactive oxygen species, which are capable to oxidize proteins. The selective removal of the latter by the 20S proteasome is considered an essential part of the cell antioxidant defense system. The aim of the present study was to investigate whether peptidase activities of rat liver proteasomes were affected by chronic (40 mg CuSO(4)/rat/daily with the drinking water for 2 weeks) and acute (20 mg/kg CuSO(4), s.c.) copper treatment. To evaluate the role of proteasome, its inhibitor MG132 was also used. The degree of copper-induced oxidative stress (OS), established by measuring lipid peroxidation, protein oxidation, and cellular glutathione level, as well as activities of antioxidant enzymes--catalase, superoxide dismutase, and gultathionine peroxidase, depended on the mode of copper administration. Chronic copper administration (mild oxidative stress) did not affect proteasome activities, whereas acute copper treatment (severe oxidative stress) caused a decline in chymotryptic- and tryptic-like activities. The treatment of copper-loaded animals with MG132 did not change copper-induced alterations in the tested indices, except an additional increase in protein oxidation and inhibition of glutathionine peroxidase activity. The results suggested that the in vivo copper-induced oxidative stress was associated with changes in the catalytic activity of proteasome.     

BODIPY-cholesterol: a new tool to visualize sterol trafficking in living cells and organisms

Analysis of sterol distribution and transport in living cells has been hampered by the lack of bright, photostable fluorescent sterol derivatives that closely resemble cholesterol. In this study, we employed atomistic simulations and experiments to characterize a cholesterol compound with fluorescent boron dipyrromethene difluoride linked to sterol carbon-24 (BODIPY-cholesterol). This probe packed in the membrane and behaved similarly to cholesterol both in normal and in cholesterol-storage disease cells and with trace amounts allowed the visualization of sterol movement in living systems. Upon injection into the yolk sac, BODIPY-cholesterol did not disturb zebrafish development and was targeted to sterol-enriched brain regions in live fish. We conclude that this new probe closely mimics the membrane partitioning and trafficking of cholesterol and, because of its excellent fluorescent properties, enables the direct monitoring of sterol movement by time-lapse imaging using trace amounts of the probe. This is, to our knowledge, the first cholesterol probe that fulfills these prerequisites.     

Genome-wide analysis of single nucleotide polymorphisms uncovers population structure in Northern Europe

Background

Genome-wide data provide a powerful tool for inferring patterns of genetic variation and structure of human populations.

Principal Findings

In this study, we analysed almost 250,000 SNPs from a total of 945 samples from Eastern and Western Finland, Sweden, Northern Germany and Great Britain complemented with HapMap data. Small but statistically significant differences were observed between the European populations (F_ST = 0.0040, p<10⁻⁴), also between Eastern and Western Finland (F_ST = 0.0032, p<10⁻³). The latter indicated the existence of a relatively strong autosomal substructure within the country, similar to that observed earlier with smaller numbers of markers. The Germans and British were less differentiated than the Swedes, Western Finns and especially the Eastern Finns who also showed other signs of genetic drift. This is likely caused by the later founding of the northern populations, together with subsequent founder and bottleneck effects, and a smaller population size. Furthermore, our data suggest a small eastern contribution among the Finns, consistent with the historical and linguistic background of the population.

Significance

Our results warn against a priori assumptions of homogeneity among Finns and other seemingly isolated populations. Thus, in association studies in such populations, additional caution for population structure may be necessary. Our results illustrate that population history is often important for patterns of genetic variation, and that the analysis of hundreds of thousands of SNPs provides high resolution also for population genetics.     

Brain serotonin transporter binding of [123I]ADAM: within-subject variation between summer and winter data

The neurotransmitter serotonin (5-HT) controls several physiological functions, and a disturbance of the 5-HT system is implicated in many psychiatric conditions. Seasonal variation has been suggested in the 5-HT system. We investigated within-subject seasonal variation in brain serotonin transporter (SERT) binding with the SERT-ligand [(123)I]ADAM and single photon emission computed tomography (SPECT) in 12 healthy individuals. No systematic variation was found in the midbrain or thalamus areas between scans done in summer and winter. Our results suggest that factors other than season are more important in causing within-subject variation of brain SERT binding between summer and winter.